LOCUS       sequence-479802-        5807 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express MAP2K2-NanoLuc Fusion Protein in Mammalian Cells under a CMV
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: MAP2K2-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2286..2798
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2886..2904)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2941..3062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2952..2971)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3006..3025
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3307..3664
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3307..3498
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3330..3350)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3515..3650
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3679..3726
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3758..4552
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3812..3831)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4422..4441
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4616..4664
                     /note="synthetic polyadenylation signal"
     rep_origin      4821..5366
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5267..5286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5483..5500
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgctg gcccggagga
     1081 agccggtgct gccggcgctc accatcaacc ctaccatcgc cgagggccca tcccctacca
     1141 gcgagggcgc ctccgaggca aacctggtgg acctgcagaa gaagctggag gagctggaac
     1201 ttgacgagca gcagaagaag cggctggaag cctttctcac ccagaaagcc aaggtcggcg
     1261 aactcaaaga cgatgacttc gaaaggatct cagagctggg cgcgggcaac ggcggggtgg
     1321 tcaccaaagt ccagcacaga ccctcgggcc tcatcatggc caggaagctg atccaccttg
     1381 agatcaagcc ggccatccgg aaccagatca tccgcgagct gcaggtcctg cacgaatgca
     1441 actcgccgta catcgtgggc ttctacgggg ccttctacag tgacggggag atcagcattt
     1501 gcatggaaca catggacggc ggctccctgg accaggtgct gaaagaggcc aagaggattc
     1561 ccgaggagat cctggggaaa gtcagcatcg cggttctccg gggcttggcg tacctccgag
     1621 agaagcacca gatcatgcac cgagatgtga agccctccaa catcctcgtg aactctagag
     1681 gggagatcaa gctgtgtgac ttcggggtga gcggccagct catagactcc atggccaact
     1741 ccttcgtggg cacgcgctcc tacatggctc cggagcggtt gcagggcaca cattactcgg
     1801 tgcagtcgga catctggagc atgggcctgt ccctggtgga gctggccgtc ggaaggtacc
     1861 ccatcccccc gcccgacgcc aaagagctgg aggccatctt tggccggccc gtggtcgacg
     1921 gggaagaagg agagcctcac agcatctcgc ctcggccgag gccccccggg cgccccgtca
     1981 gcggtcacgg gatggatagc cggcctgcca tggccatctt tgaactcctg gactatattg
     2041 tgaacgagcc acctcctaag ctgcccaacg gtgtgttcac ccccgacttc caggagtttg
     2101 tcaataaatg cctcatcaag aacccagcgg agcgggcgga cctgaagatg ctcacaaacc
     2161 acaccttcat caagcggtcc gaggtggaag aagtggattt tgccggctgg ttgtgtaaaa
     2221 ccctgcggct gaaccagccc ggcacaccca cgcgcaccgc cgtggtttct ctcggctcga
     2281 gcggcgtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     2341 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     2401 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     2461 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     2521 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     2581 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     2641 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     2701 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     2761 tgaccggctg gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agagtcgacc
     2821 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2881 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     2941 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3001 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3061 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     3121 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     3181 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3241 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     3301 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3361 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3481 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3541 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3601 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3661 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3721 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     3781 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3841 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3901 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3961 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4021 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4081 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4141 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4201 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4261 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4321 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4381 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4441 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4501 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4561 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     4621 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     4681 gatccgctca tgcgatgata agctgtcaaa catgagaatt acaacttata tcgtatgggg
     4741 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     4801 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     4861 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     4921 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     4981 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     5041 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5101 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     5161 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     5221 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     5281 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     5341 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     5401 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     5461 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     5521 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     5581 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgatgtc
     5641 cggcggtgct tttgccgtta cgcaccaccc cgtcatgaga ttacgcgttt tagcggtgcg
     5701 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     5761 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
